scholarly journals Development of the ELISA Test System for Quantitative Determination of IgG to the Varizella zoster virus in Human Serum and Assessment of its Diagnostic Efficiency

2022 ◽  
Vol 20 (6) ◽  
pp. 72-80
Author(s):  
L. N. Lukhverchyk ◽  
G. L. Alatortseva ◽  
L. N. Nesterenko ◽  
V. Y. Kabargina ◽  
V. V. Dotsenko ◽  
...  

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.

2020 ◽  
Vol 65 (11) ◽  
pp. 683-687
Author(s):  
S. G. Mardanly ◽  
A. S. Avdonina ◽  
S. G. Mamedova

A new original Russian test kit for the detection of IgG-antibodies to the causative agent of COVID-19 - coronavirus SARS-CoV-2 by the method of enzyme-linked immunosorbent assay (ELISA) on a solid-phase «ELISA-SARS-CoV-2-AT-G» has been developed. In comparative tests with similar test systems «Vitrotest® SARS-CoV-2 IgG» (Vitrotest, Ukraine) and «Anti-SARS-Cov-2 ELISA (IgG)» (EUROIMMUN AG, Germany) high diagnostic efficiency of the new test system was shown.


1988 ◽  
Vol 7 (4) ◽  
pp. 353-356 ◽  
Author(s):  
A.P. Wilkinson ◽  
D.W. Denning ◽  
M.R.A. Morgan

1 Aflatoxins are toxic, carcinogenic secondary fungal metabolites produced by certain moulds that commonly infest foods. Measurement of aflatoxins in human serum would give a direct measurement of exposure. 2 Twenty-seven serum samples from UK blood donors were found to contain aflatoxin levels not greater than 64 pmol/1 (20 pg/ml) by an enzyme-linked immunosorbent assay. 3 These findings may indicate that present UK guideline tolerances for aflatoxin in imported food are effective in limiting human exposure to toxic aflatoxins in the UK diet, though further work would be needed to confirm this. In particular, sub-populations suspected of being at higher risk may need special considerations.


2018 ◽  
Vol 69 (3) ◽  
pp. 1088
Author(s):  
A. GAVRILOVIĆ ◽  
P. GAVRILOVIĆ ◽  
S. RADOJIČIĆ ◽  
D. KRNJAIĆ

Bluetongue (BT) is a viral non-contagious disease of ruminants which is transmitted by insects of the genus Culicoides. In recent years, BT has been a serious threat to livestock and to the economies of European countries. In Serbia the disease appeared for the first time in 2001, and after a 12 year period of freedom, it broke out again in 2014. Considering the actuality of this infectious disease, especially the need for prompt and rapid diagnostics, the aim of this paper was to determine the possibility of detecting the serological response in sheep and cattle with manifested clinical signs of the disease using two different methods: double recognition enzyme-linked immunosorbent assay (sELISA) and competitive enzyme-linked immunosorbent assay (cELISA). A total of 105 blood serum samples of cattle and sheep, which had exhibited clinical signs of BT during 2014, were taken for examination from a serum bank. Out of 74 blood serum samples of sheep and 31 blood serum samples of cattle, 52 samples of sheep and 18 samples of cattle tested positive using sELISA, while 50 samples of sheep and 18 samples of cattle gave positive reactions with cELISA. The results confirm the high sensitivity of sELISA which detected 4% more seropositive sheep in comparison with cELISA. Using Cohen’s kappa statistical analysis, almost perfect agreement was determined between the results (k>0,81) obtained by cELISA and sELISA.


2016 ◽  
Vol 214 (suppl 3) ◽  
pp. S218-S221 ◽  
Author(s):  
Todd Cutts ◽  
Allen Grolla ◽  
Shane Jones ◽  
Bradley W. M. Cook ◽  
Xiangguo Qiu ◽  
...  

1987 ◽  
Vol 98 (2) ◽  
pp. 199-202 ◽  
Author(s):  
R. C. H. Lau

SUMMARYAn enzyme-linked immunosorbent assay (ELISA) incorporating the sensitive biotin-streptavidin system was developed to detect IgG antibodies to tetanus toxoid in human serum. Serum samples obtained from 557 normal persons aged 1–65 years from different areas in New Zealand were tested. The proportion of those immune ranged from 60–93% in males, and from 46–86% in females. In the 1–9 years age group 85% were immune. The indirect ELISA is suitable for serological surveys as it is simple to perform, economical and reproducible.


2012 ◽  
Vol 20 (2) ◽  
pp. 282-286 ◽  
Author(s):  
N. Ghosh ◽  
I. Tomar ◽  
H. Lukka ◽  
A. K. Goel

ABSTRACTAnthrax, caused byBacillus anthracis, is primarily a zoonotic disease. Being a public health problem also in several developing countries, its early diagnosis is very important in human cases. In this study, we describe the use of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of anti-lethal factor (anti-LF) IgG in human serum samples. A panel of 203 human serum samples consisting of 50 samples from patients with confirmed cutaneous anthrax, 93 samples from healthy controls from areas of India where anthrax is nonendemic, 44 samples from controls from an area of India where anthrax is endemic, and 16 patients with a disease confirmed not to be anthrax were evaluated with an anti-LF ELISA. The combined mean anti-LF ELISA titer for the three control groups was 0.136 ELISA unit (EU), with a 95% confidence interval (CI) of 0.120 to 0.151 EU. The observed sensitivity and specificity of the ELISA were 100% (95% CI, 92.89 to 100%) and 97.39% (95% CI, 93.44 to 99.28%), respectively, at a cutoff value of 0.375 EU, as decided by receiver operating characteristic (ROC) curve analysis. The likelihood ratio was found to be 49.98. The positive predictive value (PPV), negative predictive value (NPV), efficiency, and Youden's index (J) for reliability of the assay were 92.5%, 100%, 98.02%, and 0.97, respectively. The false-positive predictive rate and false-negative predictive rate of the assay were 2.61% and 0%. The assay could be a very useful tool for early diagnosis of cutaneous anthrax cases, as antibodies against LF appear much earlier than those against other anthrax toxins in human serum samples.


2001 ◽  
Vol 69 (5) ◽  
pp. 3224-3231 ◽  
Author(s):  
Fang Ting Liang ◽  
Lisa C. Bowers ◽  
Mario T. Philipp

ABSTRACT VlsE, the variable surface antigen of Borrelia burgdorferi, contains two invariable domains located at the amino and carboxyl terminal ends, respectively, and a central variable domain. In this study, both immunogenicity and antigenic conservation of the C-terminal invariable domain were assessed. Mouse antiserum to a 51-mer synthetic peptide (Ct) which reproduced the entire sequence of the C-terminal invariable domain of VlsE from B. burgdorferi strain B31 was reacted on immunoblots with whole-cell lysates extracted from spirochetes of 12 strains from the B. burgdorferi sensu lato species complex. The antiserum recognized only VlsE from strain B31, indicating that epitopes of this domain differed among these strains. When Ct was used as enzyme-linked immunosorbent assay (ELISA) antigen, all of the seven monkeys and six mice that were infected with B31 spirochetes produced a strong antibody response to this peptide, indicating that the C-terminal invariable domain is immunodominant. None of 12 monkeys and only 11 of 26 mice that were infected with strains other than B31 produced a detectable anti-Ct response, indicating a limited antigenic conservation of this domain among these strains. Twenty-six of 33 dogs that were experimentally infected by tick inoculation were positive by the Ct ELISA, while only 5 of 18 serum samples from dogs clinically diagnosed with Lyme disease contained detectable anti-Ct antibody. Fifty-seven of 64 serum specimens that were collected from American patients with Lyme disease were positive by the Ct ELISA, while only 12 of 21 European samples contained detectable anti-Ct antibody. In contrast, antibody to the more conserved invariable region IR6 of VlsE was present in all of these dog and human serum samples.


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